Chinese Hamster Cells In Culture Research Articles

Demographic Approaches to the Study of Aging on Cell Cultures

Aging organisms die out in accordance with the “Gompertz law,” i.e., the probability of their death increases with age. Survival curve construction is the main tool for gerontologists to study aging and test antiaging drugs. The analysis of survival curves includes obtaining some indices characterizing aging of the population, for example, the average and maximum lifespan, the mortality rate, and the aging rate. Testing geroprotectors can be correctly performed only by obtaining such curves. The dying out of stationary cell populations—bacteria, yeast, and mammalian cell cultures—also occurs in accordance with the Gompertz equation. In this regard, it is reasonable to use the construction of survival curves and their analysis to study the “aging” of non-subcultured cell cultures and testing anti-aging drugs on them. We used this approach in our experiments, due to which we were able to detect the positive anti-aging effect of the Quinton Marine Plasma on stationary phase aging culture of Chinese hamster cells.

Studies into the Effect of “Mild” Uncoupling with 2,4-Dinitrophenol on the Growth of Chinese Hamster Cell Culture and Its Subsequent Dying out in the Stationary Phase

Partial uncoupling of the processes of oxidative phosphorylation and energy storage in the form of ATP (“mild” uncoupling) helps reduce the production of reactive oxygen species and can also mimic the effect of calorie restriction. A number of studies have shown that uncouplers, such as 2,4-dinitrophenol (DNP), affect the lifespan of Drosophila, yeast, mice, and rats as well as the manifestation of “age-related” changes in cultures of mammalian and human cells undergoing replicative senescence. This paper is devoted to studying the effect of DNP on the growth and subsequent dying out of “stationary phase aging” Chinese hamster cells. Using the method for evaluating the colony-forming efficiency of cells, the maximum permissible concentration was selected, 5 ×10–5 M, in which the substance presumably induces “mild” uncoupling and does not inhibit cell proliferation. At higher concentrations, DNP has a cytotoxic effect on the studied cell culture. Under the influence of DNP in the potentially “mild” uncoupling concentration (5.6 × 10–7 M), the kinetics of cell growth and dying out does not change, and the lifespan of the cell culture does not increase. This effect may be due to the type of cells studied. In addition, there is a probability that the optimal concentration lies in the range from 5 ×10–7 to 5 × 10–5 M or even lower than 5 × 10–7 M.

Chromosome-breaking capacity of tepa and analogues in Vicia faba and Chinese hamster cells

The insect chemosterilant tris(l-aziridinyl)phosphine oxide, tepa, with three aziridinyl groups, six tepa analogues with two aziridinyl groups and carbon chains of varying length, one monoaziridinyl analogue of tepa, and hempa with no aziridinyl group have been tested for chromosome-breaking ability in root tips of Vicia faba and in cell cultures of Chinese hamster. The difunctional analogues proved to be as active as tepa in both materials; the monofunctional analogue was less effective and hempa produced no effect. The hamster cells proved to be 23 times more sensitive than the bean roots to the chromosome-breaking effect of tepa. In the hamster cells the chromatid break was the most frequent type of aberration. Incomplete chromosomal aberrations were more numerous in the hamster cells than in the root tips. Combined treatments with tepa and [3H]-thymidine were performed to determine the sensitivity of the various phases of the mitotic cycle to the chromosome-breaking effect in the two materials. Treatment with tepa induced only chromatid-type aberrations. Chromosomal aberrations at the first mitosis (T1) were observed only in cells treated with tepa while in the DNA synthesis (S) and pre-synthetic (G1) periods, but not in the post-synthetic (G2) period. In the second mitosis (T2), G2 cells which were visibly undamaged at T1 mitosis, contained chromatid-type aberrations. The frequency of aberrations observed in T2 mitosis was higher than in T1 mitosis. Thus, although tepa produces lesions in chromosomes independently of cell phase at the time of treatment, these lesions apparently require a period of DNA synthesis before they are converted to chromatid aberrations observable in the following mitosis.

Induction of apoptosis in populations of mammalian cells in vitro under the influence of lectins

A study of the cytogenetic action of lectins that differ in terms of origin and hydrocarbon specification is presented. In a culture of Chinese hamster cells it is shown that at high concentrations (20 μg/ml), all lectins are capable of inducing an increase in the level of apoptosis two days after treatment with a high degree of reliability. In the case of perch roe lectin, such an increase occurs both immediately after treatment as well as two days later. It is found that, unlike lentil and perch roe lectin, in a culture of human cells black elder lectin does not have any effect on the incidence of apoptotic cells. A trend towards stimulation of proliferation of human cells under the influence of perch roe lectin at low concentrations (0.2 μg/ml) is discovered.

Poly(ADP-Ribose)-polymerase-1 and aging: Experimental study of possible relationship on stationary cell cultures

Experiments on "stationary aging" cultures of B11dii-FAF28 Chinese hamster cells showed that contact inhibition of cell growth and further culturing of cells in the stationary phase led to continuous inhibition of enzymes realizing poly-ADP-ribosylation of chromatin proteins. Cell density in the monolayer and percentage of damaged cells detected by trypan blue staining decreases during this process.

Instability of chromosome structure in cancer cells increases exponentially with degrees of aneuploidy

Structurally altered or marker chromosomes are the cytogenetic hallmarks of cancer cells, but their origins are still debated. Here we propose that aneuploidy, which is ubiquitous in cancer and inevitably unbalances thousands of synergistic genes, destabilizes the structure of chromosomes by catalyzing DNA breaks. Aneuploidy catalyzes such breaks by unbalancing teams of enzymes, which synthesize and maintain DNA and nucleotide pools, and even unbalancing histones via the corresponding genes. DNA breaks then initiate deletions, amplifications, and intra- and interchromosomal rearrangements. Our hypothesis predicts that the rate at which chromosomes are altered is proportional to the degree of aneuploidy: the more abnormal the number and balance of chromosomes, the higher the rate of structural alterations. To test this prediction, we have determined the rates at which clonal cultures of diploid and aneuploid Chinese hamster cells generate new, and thus nonclonal, structurally altered chromosomes per mitosis. Based on about 20 metaphases, the number of new, structurally altered chromosomes was 0 per diploid, 0–0.23 per near-diploid, 0.2–1.4 per hypotriploid, 3.25–4.8 per hypertriploid, and 0.4 per near-tetraploid cell. Thus, instability of chromosome structure increases exponentially with the deviation of ploidy from the normal diploid and tetraploid balances. The particular chromosomes engaged in aneuploidy also affected the rates of chromosome alteration, particularly at low aneuploidy indices. We conclude that aneuploidy is sufficient to cause structural instability of chromosomes. Further, we suggest that certain structurally altered chromosomes encode cancer-specific phenotypes that cannot be generated by unbalancing intact chromosomes. We also extend the evidence for aneuploidy causing numerical instability of chromosomes autocatalytically, and adduce evidence that aneuploidy can cause the many gene mutations of cancer cells that have been attributed to various mutator genes.

Cell studies of the DNA bis-intercalator Delta-Delta [mu-C4(cpdppz)(2)-(phen)(4)Ru(2)](4+): toxic effects and properties as a light emitting DNA probe in V79 Chinese hamster cells.

Coordination complexes of type [Ru(L)(3)](2+), where L is a nitrogen-containing aromatic bidentate ligand, can often be photolytically reduced, making them useful in studies of DNA- or protein-mediated electron transfer and in artifical photosynthesis model systems. Upon binding to DNA some Ru(L) complexes have been found to display strongly increased fluorescence compared with when free in solution, making those compounds interesting to test as DNA probes. Thus, they are becoming widely used in the chemistry community. Here, asynchronous cultures of V79 Chinese hamster cells were exposed to the DNA bis-intercalator Delta-Delta [mu-C4(cpdppz)(2)-(phen)(4)Ru(2)](4+) at 10(-10)-10(-4) M. The extraordinarily strong binding of the compound to DNA was the reason for testing its possible interference with DNA metabolism in intact mammalian cells. Exposure for 1 h to 10(-10)-10(-4) M did not significantly decrease DNA synthesis. Cells exposed to 10(-5) M for 27 h showed no staining of the nucleus, while DNA was stained in cells electroporated in the presence of the compound. However, the Ru dimer was probably taken up by pinocytosis, because numerous minute precipitates could be observed in the cytoplasm. Treatment for 24 h at concentrations of 10(-10)-10(-5) M did not inhibit growth, as indicated by cell density and mitotic activity. Neither did it affect chromosomal arrangements during mitosis. However, at 10(-4) M the density of cultures was reduced by approximately 45% and apoptotic cells were frequent, as opposed to mitoses. We also investigated the properties of the Ru dimer as a fluorescent DNA stain. The compound appears attractive as a red DNA stain when broad excitation in the visible range is desirable and extremely low background staining is essential. The low toxicity of the compound is a favourable trait in this context.

Final Report on the Safety Assessment of Cocoyl Sarcosine, Lauroyl Sarcosine, Myristoyl Sarcosine, Oleoyl Sarcosine, Stearoyl Sarcosine, Sodium Cocoyl Sarcosinate, Sodium Lauroyl Sarcosinate, Sodium Myristoyl Sarcosinate, Ammonium Cocoyl Sarcosinate, and Ammonium Lauroyl Sarcosinate

This safety assessment addresses cosmetic ingredients that are N-acyl derivatives of sarcosine and are generally referred to as acyl sarcosines, and those that are salts, known generally as acyl sar-cosinates. Previous assessments have addressed the safety of each of the fatty acids that appear in these acyl sarcosines and sarcosinates (Coconut Acid, Oleic Acid, Lauric Acid, and Myristic Acid). In each case the fatty acid was either safe for use or safe as used in cosmetic formulations. Acyl sarcosines are considered modified fatty acids with greater solubility and increased acidity of the carboxylic acid group compared to the parent fatty acid. They are used in a large number of cosmetic formulations as hair-conditioning agents and surfactant-cleansing agents. In soaps, concentrations are reported to be as high as 12.9%. These ingredients have low oral toxicity in rats. Although cytotoxic to Chinese hamster cells in culture, acyl sarcosines and sarcosinates are not mutagenic in those cells, nor in bacterial cells in culture. Carcinogenicity data were not available. These ingredients are nonirritating and nonsen-sitizing to animal and human skin, although they can enhance the penetration of other ingredients through the skin. For that reason, caution should be exhibited in formulating cosmetic products that contain these ingredients in combination with other ingredients whose safety is based on their lack of absorption or where dermal absorption is a concern (e.g., HC Yellow No. 4, Disperse Yellow 3). Because sarcosine can be nitrosated to form N-nitrososarcosine, a known animal carcinogen, these ingredients should not be used in cosmetic products in which N-nitroso compounds may be formed. With the above caveat, and based on the available data, it was concluded that these acyl sarcosines and sarcosinates are safe as used in rinse-off products. They may be safely used in leave-on products at concentrations up to 5%, the highest concentration tested in clinical irritation and sensitization studies. Oleoyl Sarcosine is used as a corrosion inhibitor in some aerosol products, at extremely low concentrations. In this circumstance, the ingredient is not being used as a cosmetic ingredient and this report is not intended to limit that use. Because of the absence of data on inhalation toxicity, however, it was concluded that the available data were not sufficient to support the safety of acyl sarcosines and sarcosinates as cosmetic ingredients in products where they are likely to be inhaled.

Final Report on the Safety Assessment of Cocoyl Sarcosine, Lauroyl Sarcosine, Myristoyl Sarcosine, Oleoyl Sarcosine, Stearoyl Sarcosine, Sodium Cocoyl Sarcosinate, Sodium Lauroyl Sarcosinate, Sodium Myristoyl Sarcosinate, Ammonium Cocoyl Sarcosinate, and Ammonium Lauroyl Sarcosinate

This safety assessment addresses cosmetic ingredients that are N-acyl derivatives of sarcosine and are generally referred to as acyl sarcosines, and those that are salts, known generally as acyl sar-cosinates. Previous assessments have addressed the safety of each of the fatty acids that appear in these acyl sarcosines and sarcosinates (Coconut Acid, Oleic Acid, Lauric Acid, and Myristic Acid). In each case the fatty acid was either safe for use or safe as used in cosmetic formulations. Acyl sarcosines are considered modified fatty acids with greater solubility and increased acidity of the carboxylic acid group compared to the parent fatty acid. They are used in a large number of cosmetic formulations as hair-conditioning agents and surfactant-cleansing agents. In soaps, concentrations are reported to be as high as 12.9%. These ingredients have low oral toxicity in rats. Although cytotoxic to Chinese hamster cells in culture, acyl sarcosines and sarcosinates are not mutagenic in those cells, nor in bacterial cells in culture. Carcinogenicity data were not available. These ingredients are nonirritating and nonsen-sitizing to animal and human skin, although they can enhance the penetration of other ingredients through the skin. For that reason, caution should be exhibited in formulating cosmetic products that contain these ingredients in combination with other ingredients whose safety is based on their lack of absorption or where dermal absorption is a concern (e.g., HC Yellow No. 4, Disperse Yellow 3). Because sarcosine can be nitrosated to form N-nitrososarcosine, a known animal carcinogen, these ingredients should not be used in cosmetic products in which N-nitroso compounds may be formed. With the above caveat, and based on the available data, it was concluded that these acyl sarcosines and sarcosinates are safe as used in rinse-off products. They may be safely used in leave-on products at concentrations up to 5%, the highest concentration tested in clinical irritation and sensitization studies. Oleoyl Sarcosine is used as a corrosion inhibitor in some aerosol products, at extremely low concentrations. In this circumstance, the ingredient is not being used as a cosmetic ingredient and this report is not intended to limit that use. Because of the absence of data on inhalation toxicity, however, it was concluded that the available data were not sufficient to support the safety of acyl sarcosines and sarcosinates as cosmetic ingredients in products where they are likely to be inhaled.

Inhibition of mitotic-specific histone phosphorylation by sodium arsenite

Synchronized cultures of Chinese hamster cells (line CHO) were used to measure the effects of 10 μ m sodium arsenite on histone phosphorylation. This treatment caused cell proliferation to be temporarily arrested, after which the cells spontaneously resumed cell proliferation in a radiomimetic manner. Immediately following treatment, it was found that sodium arsenite affected only mitotic-specific H1 and H3 phosphorylations. Neither interphase, nor mitotic, H2A and H4 phosphorylations were affected, nor was interphase H1 phosphorylation affected. The phosphorylation of H1 was inhibited only in mitosis, reducing H1 phosphorylation to 38.1% of control levels, which was the level of interphase H1 phosphorylation. The phosphorylation of both H3 variants was inhibited in mitosis, the less hydrophobic H3 to 19% and the more hydrophobic H3 to 24% of control levels. These results suggest that sodium arsenite may inhibit cell proliferation by interfering with the cyclin B/p34 cdc2 histone kinase activity which is thought to play a key role in regulating the cell cycle. It has been proposed that H1 and H3 phosphorylations play a role in restructuring interphase chromatin into metaphase chromosomes. Interference in this process by sodium arsenite may lead to structurally damaged chromosomes, resulting in the increased cancer risks known to be produced by arsenic exposure from the environment.

Metabolism of phenacetin in V79 Chinese hamster cell cultures expressing rat liver cytochrome P4501A2 compared to isolated rat hepatocytes

In a genetically engineered V79 cell line (XEMd-MZ) expressing rat cytochrome P4501A2 the activity of phenacetin- O-deethylase was determined and compared with freshly isolated rat hepatocytes. In the V79 cells the apparent K m , was 0.99 μM (N = 4), compared to the high affinity K m (0.23 μM, N = 4) found in freshly isolated rat hepatocytes, where as the V max found in the XEMd-MZ cells (14.90 pmol/min/10 6 cells) corresponded to the V max for the high affinity site of the hepatocytes (18.2 pmol/min/10 −6 cells). The metabolic capacity of phenacetin was quantitatively comparable to that of freshly isolated hepatocytes on a cell to cell basis.

Radioprotective effect of N-acetylcysteine in vitro using the induction of DNA breaks as end-point.

Cultures of Chinese hamster cells were treated with N-acetylcysteine (NAC) and irradiated under hypoxic or aerobic conditions. Protection factors (PFs) were calculated using the yield of DNA breaks as a measure of the radiation response. With NAC up to 50 mmol dm-3, the PF values were smaller for aerobically- than for hypoxically-irradiated cells. In contrast with NAC concentrations > 100 mmol dm-3, the PF values for aerobically-irradiated cells were larger than those for hypoxic cells. This reversal is in agreement with the predictions of a modified competition model, the 'X-model'. The value of X (the ratio of reparable to non-reparable damage) was calculated to be close to 5 for the NAC-treated cells, i.e. about 18% of the radiation damage was not reparable. The increase of the total non-protein bound thiols (NPSH) and GSH content after NAC treatment was measured both in whole cells and isolated nuclei. Measurement of the cellular NPSH content showed a continuous increase with increasing NAC concentrations in the medium, independent of treatment time. In contrast, the amount of intracellular and intranuclear GSH increased with prolonged incubation time with NAC. No quantitative relationship was found between PF values and cellular NPSH or GSH content.

N-Methyl- N′-nitro- N-nitrosoguanidine-induced light emission in Chinese hamster cell cultures: correlation with enhancement of chromosomal aberrations

N-Methyl- N′-nitro- N-nitrosoguanidine (MNNG) was found to induce an ultraweak photon emission in cultures of Chinese hamster fibroblasts (CHL). Measurements suggest that the light emission is due to a reaction between MNNG and cellular metabolites. The light emission depended on the concentration of MNNG and was oxygen-dependent, disappearing in a nitrogen atmosphere. Superoxide dismutase (SOD) or sodium azide decreased the emission intensity. The production of chromosomal aberrations in CHL by MNNG was correlated with the light emission intensity and was inhibited in the presence of SOD.

Protection by the fluoride ion against phthalocyanine-induced photodynamic killing of Chinese hamster cells.

When a dilute F- solution was added to a culture of Chinese hamster cells that had been preincubated with an aluminium phthalocyanine sensitizer derived from AlPcCl, the photosensitivity of the cells was markedly reduced compared to control cells not treated with F-. Under the same treatment conditions, the reduction in [3H]thymidine incorporation into cellular DNA caused by light and this sensitizer and the production of DNA-protein crosslinks caused by light and this sensitizer were also inhibited by F-. In contrast, the killing of Chinese hamster cells, the reduction of thymidine incorporation by the cells, and the production of DNA-protein crosslinks in the cells caused by the combination of light and either Photofrin II or the silicon phthalocyanine HOSiPcOSi(CH3)2(CH2)3-N(CH3)2 were not inhibited by F-. We conclude that the aluminium phthalocyanine sensitizer used is largely or completely AlPc(OH)(H2O), that it is converted to a fluoro complex by F-, and that this compound probably is a less efficient generator of photochemical damage at a critical cellular target(s) than is AlPc(OH)(H2O). The inhibition of thymidine incorporation and DNA-protein crosslink formation indicates that the effects of F- can be expressed at intracellular sites. It is further concluded that the silicon phthalocyanine sensitizer and Photofrin II do not interact significantly with F-.

Induction of chromosomal aberrations and SCE by chloramphenicol

The induction of chromosomal aberrations and sister-chromatid exchanges (SCE) was studied in human lymphocyte cultures treated with chloramphenicol (CAP), an antimicrobiol agent acting by inhibiting protein synthesis. Moreover chromosomal aberrations and sister-chromatid exchanges were studied in bone marrow cells of treated mice and in Chinese hamster cell cultures (V79) respectively. While no aberrations were induced by short treatments in human lymphocytes exposed in G 1 and G 2 phases, high frequencies of aberrations, exclusively of the chromatid type, were induced when the drug was administered during a whole cell cycle. Aberrant metaphases were detected only at the end and a few hours after the end of treatment; at later times aberrant cells reached control values. Doses producing aberrations only slightly increased SCE both in human lymphocytes and in V79 cells. In mouse bone marrow cells CAP induced a high mitotic delay and few structural aberrationsl intrachromosomal vacuoles were observed.

Synthesis and photocytotoxicity of some new substituted phthalocyanines.

Some new, ring-substituted phthalocyanines have been synthesized and underwent preliminary testing for photodynamic activity using the colony forming ability of Chinese hamster cells in culture as an endpoint. Using 4-(3-N,N-diethylaminopropyl)phthalonitrile as a precursor, the previously unknown metal-free 2,9,16,23-tetra-(3-N,N-diethylaminopropyl)phthalocyanine was prepared and converted to its zinc (II) and its cationic water-soluble 2,9,16,23-tetramethylammonium zinc (II) iodide derivatives. Other new phthalocyanine derivatives tested, include a 2,9,16,23-tetra(2-hydroxymethyl-2-methylbutoxy)phthalocyaninato zinc (II) and 2,3,9,10,16,17,23,24-octahydroxyphthalocyaninato zinc (II) derivatives. Boron tribromide cleavage of the newly prepared and fully characterized 2,3,9,10,16,17,23,24-octamethoxyphthalocyanine gave the unstable metal-free octahydroxyphthalocyanine, which could only be characterized by ultraviolet-visible, NMR and IR spectroscopy.

Protective role of potentially lethal damage repair in the neoplastic transformation of Balb/c 3T3 cells treated with N-methyl-N′-nitro-N-nitrosoguanidine

To determine the role of repair of potentially lethal damage (PLD) in the initiation process of neoplastic transformation, BALB/c 3T3 cells treated with N-methyl-N′-nitro-N-nitrosoguanidine (MNNG) were temporarily exposed to conditioned medium obtained from density-inhibited Chinese hamster cell cultures, as a post-treatment for the induction of PLD repair. With or without this exposure, cell survival and transformation frequencies were simultaneously determined by colony-formation and focus-formation assays, respectively. Temporary exposure to conditioned medium resulted in a 20–30% increase in cell survival compared with no exposure. Post-treatment with conditioned medium resulted in a 60–65% reduction in transformation frequencies. At the molecular level, the repair of MNNG-induced single-strand breaks of DNA occurred much more rapidly in condition medium. These data suggest that PLD repair DNA replication. Thus, PLD repair appears to be preventive against neoplastic fixation in initiation of neoplastic development.

Induction of chromosomal aberrations in active oxygen-generating systems: Studies on soft X-rays in Chinese hamster cells in culture

Induction of chromosomal aberrations in active oxygen-generating systems: Studies on soft X-rays in Chinese hamster cells in culture

Methotrexate-poly(lysine) as a selective agent for mutants of Chinese hamster ovary cells defective in endocytosis.

Methotrexate (MTX) covalently linked to poly(L-lysine) [poly(Lys)] enters cells by endocytosis, is degraded in lysosomes and, upon liberation of small molecular methotrexate, is cytocidal to Chinese hamster cells in culture. This drug conjugate was used to select mutants resistant to MTX-poly(Lys), which were examined for defects in endocytosis. Two mutants resistant to MYX-poly(Lys) and sensitive to free MTX, MPL 3-4 and MPL 2-5, internalized the conjugate in normal fashion, but had a decreased ability to degrade it to small molecular drug. The magnitude of this defect in the two mutants correlated with their level of resistance. In addition, both mutants were cross resistant to diphtheria toxin and modeccin and hypersensitive to ricin. While MPL 3-4 internalized MTX-poly(Lys) and inulin normally, it showed decreased endocytosis via the mannose-6-phosphate receptor and decreased uptake of 125I-alpha-2 macroglobulin. Acidification of subcellular fractions was measured using the partitioning of acridine orange. In MPL 3-4, the ATP-driven acidification of the endosome-containing cell fractions was slightly decreased (80% of controls), while acidification of the heavy lysosome-containing fraction was normal. Complementation analysis using hybrids of MPL 3-4 x MPL 2-5 indicated that the mutations occurred at the same gene, but were expressed with different severity. This genotype is identical to that of the End 2 mutants described by Roff et al. (1986). Thus, surprisingly, mutants with identical genotypes were isolated independently by totally different selection procedures.

Comparison of the mutagenicity of amsacrine with that of a new clinical analogue, CI-921

The mutagenicity of CI-921, the 4-methyl-5-( N-methyl)carboxamide derivative of the clinical antileukaemia agent, amsacrine, has been assessed using both bacterial and mammalian cells. CI-921 is distinguished from amsacrine in its high activity against some experimental tumours and is currently undergoing phase I clinical trial. Like 9-aminoacridine and amsacrine, CI-921 is mutagenic to the Salmonella typhimurium frameshift tester strain TA1537, but shows no sign of inducing base pair changes in strain TA100. In Chinese hamster cell culture, however, it differs from 9-aminoacridine in causing extensive chromosomal aberrations and an increase in mutations at the hypoxanthine—guanine phosphoribosyltransferase locus. It induces the formation of tightly packed and multilayered colonies in treated cultures of C 3H/10T 1 2 cells, but its action differs from that of benzo[ a]pyrene, which induces type III fibroblastic multilayered colonies. Side-by-side comparison of the mutagenic properties of CI-921 and amsacrine showed no substantial differences at similar toxicity, suggesting that the increased lipophilicity and DNA-binding affinity of CI-921, which are thought to contribute to its increased antitumour activity, do not concomitantly increase the efficiency of in vitro mutagenesis or cell transformation.